Therapeutic compounds

ABSTRACT

A compound having a substituted five or six-membered carbocycle or heterocycle directly bonded to a substituted aryl or heteroaryl ring, wherein said compound has an EC 50  value of 20 nM or less at the prostaglandin EP 2  receptor according to the cAMP assay. 
     Methods, compositions, and medicaments related thereto are also disclosed.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of U.S. patent applicationSer. No. 11/009,298, filed on Dec. 10, 2004 now U.S. Pat. No. 7,091,231,the disclosure of which is expressly incorporated by reference herein.

DESCRIPTION OF THE INVENTION

A compound having a substituted five or six-membered carbocycle orheterocycle directly bonded to a substituted aryl or heteroaryl ring,wherein said compound has an EC₅₀ value of 20 nM or less in HEK-EBNAcells expressing a PG EP2 receptor subtype according to the cAMP assayis disclosed herein.

In one embodiment, the compound reduces intraocular pressure of a beagledog by at least 4 mmHg when administered topically in a liquidcomposition to an eye of said dog, wherein compound has a concentrationof about 0.3% (w/v) or less in said composition.

Intraocular pressure studies in dogs involved pneumatonometry performedin conscious, Beagle dogs of both sexes (10-15 kg). The animals remainedconscious throughout the study and were gently restrained by hand. Drugswere administered topically to one eye as a 25 μL volume drop, the othereye received 25 μL vehicle (0.1% (w/v) polysorbate 80; 10 mM TRIS) as acontrol. Proparacaine (0.1% w/v) was used for corneal anesthesia duringtonometry. Intraocular pressure was determined just before drugadministration and at 2, 4 and 6 hr thereafter on each day of the 5 daystudy. Drug was administered immediately after the first IOP reading.

In another embodiment, the compound reduces intraocular pressure of ahuman with glaucoma or elevated intraocular pressure by at least 5 mmHgwhen administered topically in a liquid composition to an eye of saidhuman, wherein compound has a concentration of about 0.3% (w/v) or lessin said composition.

“The cAMP assay” is:

A 384-well drug plate was prepared to contain 6 test compounds, PGE2 andcAMP in 16 serial dilutions in triplicate, using a Biomek station.HEK-EBNA cells expressing a target PG receptor subtype (EP2 or EP4) weresuspended in a stimulation buffer (HBSS, 0.1% BSA, 0.5 mM IBMX and 5 mMHEPES, pH 7.4) in a density of 10⁴ cells/5 μl. The reaction wasinitiated by mixing 5 μL drug dilutions with 5 μl of HEK-EBNA cells in awell, carried out for 30 min at room temperature, and followed by theaddition of 5 μl anti-cAMP acceptor beads in the control buffer withTween-20 (25 mM NaCl, 0.03% Tween-20, 5 mM HEPES, pH7.4). After 30 minin the dark at room temperature, the mixtures were incubated with 15 μlbiotinylated-cAMP/strepavidin donor beads in Lysis/Detection buffer(0.1% BSA, 0.3% Tween-20 and 5 mM HEPES, pH7.4) for 45 min at the roomtemperature. Fluorescence changes were read using a Fusion-alpha HTmicroplate reader.

Reduction of intraocular pressure of a beagle dog by at least 4 mmHg isdetermined by the procedure described herein.

Reduction of intraocular pressure in a human with glaucoma or elevatedintraocular pressure by at least 5 mmHg is done on a person whoseintraocular pressure is elevated more than 5 mmHg above normal.

Another embodiment is a compound of the formula

or a pharmaceutically acceptable salt thereof, or a prodrug thereof, ora tautomer thereof;

-   wherein a dashed line represents the presence or absence of a bond,    provided that an atom does not have a double bond to two different    ligands;-   Y is an organic acid functional group, or an amide or ester thereof    comprising up to 14 carbon atoms; or Y is hydroxymethyl or an ether    thereof comprising up to 14 carbon atoms; or Y is a tetrazolyl    functional group;-   A is —(CH₂)₆—, cis —CH₂CH═CH—(CH₂)₃—, or —CH₂C≡C—(CH₂)₃—, wherein 1    or 2 carbon atoms may be replaced by S or O; or A is    —(CH₂)_(m)—Ar—(CH₂)_(o)— wherein Ar is interarylene or    heterointerarylene, the sum of m and o is 1, 2, 3, or 4, and wherein    one CH₂ may be replaced by S or O;-   Z¹, Z², Z³, Z⁴, Z⁵, and Z⁶ are independently CH₂, CH, C, NH, or N;    provided that at least one of Z¹, Z², Z³, Z⁴, Z⁵, and Z⁶ is not NH    or N;-   J¹, J², J³, J⁴ are independently hydrogen; F; Cl, Br; I; O; OH; S;    SH; O-alkyl having 1, 2, 3, 4, 5 or 6 carbon atoms;-   alkyl having 1, 2, 3, 4, 5, or 6 carbon atoms; or CF₃; and-   B is substituted aryl or substituted heteroaryl.

Another embodiment is a compound comprising

or a pharmaceutically acceptable salt or a prodrug thereof,

-   wherein a dashed line represents the presence or absence of a    covalent bond;-   Y is a carboxylic acid, sulfonic acid, or phosphonic acid; or an    amide or ester thereof comprising from 0 to 12 carbon atoms; or Y is    a hydroxymethyl, or tetrazolyl functional group;-   A is —(CH₂)₆—, cis —CH₂CH═CH—(CH₂)₃—, or —CH₂C≡C—(CH₂)₃—, wherein 1    or 2 carbon atoms may be substituted with S or O; or A is    —(CH₂)_(m)—Ar—(CH₂)_(o)— wherein Ar is substituted or unsubstituted    phenyl or monocyclic heteroaryl, the sum of m and o is from 1 to 4,    and wherein one CH₂ may be substituted with S or O;-   X is C═O, CHF, CF₂, CHCl, or CHOH; wherein if X is CHOH, then OH is    in the β-configuration;-   R is a hydrocarbyl or a hydroxyhydrocarbyl moiety comprising from 1    to 12 carbon atoms;-   D is independently a moiety comprising from 1 to 6 non-hydrogen    atoms; and n is an integer from 0 to 4.

A compound comprising a prostaglandin EP₂ selective agonist wherein theω-chain comprises a substituted phenyl, wherein at least one substituentconsists of hydrocarbyl or non-linear hydroxyhydrocarbyl, is alsodisclosed herein.

Several of the carbon atoms on these compounds are chiral centers. Whilenot intending to limit the scope of the invention in any way, or bebound in any way by theory, it is believed that many compounds andpharmaceutically active salts or prodrugs thereof having thestereochemistry shown below are particularly useful.

A person of ordinary skill in the art understands the meaning of thestereochemistry associated with the hatched wedge/solid wedge structuralfeatures. For example, an introductory organic chemistry textbook(Francis A. Carey, Organic Chemistry, New York: McGraw-Hill Book Company1987, p. 63) states “a wedge indicates a bond coming from the plane ofthe paper toward the viewer” and the hatched wedge, indicated as a“dashed line”, “represents a bond receding from the viewer.”

However, it is also advantageous if one or more of the bonds has theindicated stereochemistry, while the stereochemistry of other bonds tochiral centers may vary. Thus, while not intending to limit the scope ofthe invention in any way, compounds comprising

and the like, and pharmaceutically acceptable salts and prodrugsthereof, are particularly useful in the context disclosed herein.

A “pharmaceutically acceptable salt” is any salt that retains theactivity of the parent compound and does not impart any additionaldeleterious or untoward effects on the subject to which it isadministered and in the context in which it is administered compared tothe parent compound. A pharmaceutically acceptable salt also refers toany salt which may form in vivo as a result of administration of anacid, another salt, or a prodrug which is converted into an acid orsalt.

Pharmaceutically acceptable salts of acidic functional groups may bederived from organic or inorganic bases. The salt may comprise a mono orpolyvalent ion. Of particular interest are the inorganic ions, lithium,sodium, potassium, calcium, and magnesium. Organic salts may be madewith amines, particularly ammonium salts such as mono-, di- and trialkylamines or ethanol amines. Salts may also be formed with caffeine,tromethamine and similar molecules. Hydrochloric acid or some otherpharmaceutically acceptable acid may form a salt with a compound thatincludes a basic group, such as an amine or a pyridine ring.

A “prodrug” is a compound which is converted to a therapeutically activecompound after administration, and the term should be interpreted asbroadly herein as is generally understood in the art. While notintending to limit the scope of the invention, conversion may occur byhydrolysis of an ester group or some other biologically labile group.Generally, but not necessarily, a prodrug is inactive or less activethan the therapeutically active compound to which it is converted.

Y is a carboxylic acid, sulfonic acid, or phosphonic acid functionalgroup; or an amide or ester thereof comprising from 0 to 12 carbonatoms; or Y is a hydroxymethyl, or tetrazolyl functional group. Thus,while not intending to limit the scope of the invention in any way, incertain compounds Y is a carboxylic acid, sulfonic acid, or phosphonicacid functional group, i.e. one of the structures shown below.

Salts of any of these acids of any pharmaceutically acceptable form mayalso be present.

Additionally, an amide or ester of one of the organic acids shown abovecomprising from 0 to 12 carbon atoms is also contemplated. In an ester,a hydrocarbyl moiety replaces a hydrogen of an acid such as in acarboxylic acid ester, e.g. CO₂R³. In an amide, an amine group replacesan OH of the acid. An amine is a moiety having a central nitrogen whichhas exactly three bonds to C or H. Examples of amides include CON(R³)₂,CON(OR³)R³, CON(CH₂CH₂OH)₂, and CONH(CH₂CH₂OH). Moieties such asCONHSO₂R³ are also amides of the carboxylic acid notwithstanding thefact that they may also be considered to be amides of the sulfonic acidR³—SO₃H.

Finally, while not intending to limit the scope of the invention in anyway, Y may also be a hydroxymethyl, or a tetrazolyl functional group,i.e. compounds having a structure such as one of those shown below.

When R³ is hydrogen, the tetrazolyl functional group has two tautomericforms, which can rapidly interconvert in aqueous or biological media,and are thus equivalent to one another. These tautomers are shown below.

Additionally, if R³ is C₁-C₆ alkyl, phenyl, or biphenyl, other isomericforms of the tetrazolyl functional group such as the one shown below arealso possible, all of these are considered to be within the scope of theterm “tetrazolyl.”

While not intending to limit the scope of the invention in any way, inone embodiment, Y is selected from the group consisting of CO₂(R³),CON(R³)₂, CON(OR³)R³, CON(CH₂CH₂OH)₂, CONH(CH₂CH₂OH), CH₂OH, P(O)(OH)₂,CONHSO₂R³, SO₂N(R³)₂, SO₂NHR³, and tetrazolyl-R³; wherein R³ isindependently H, C₁-C₆ alkyl, phenyl, or biphenyl.

In relation to the identity of A disclosed in the chemical structurespresented herein, in the broadest sense, A is —(CH₂)₆—, cis—CH₂CH═CH—(CH₂)₃—, or —CH₂C≡C—(CH₂)₃—, wherein 1 or 2 carbon atoms maybe substituted with S or O; or A is —(CH₂)_(m)—Ar—(CH₂)_(o)— wherein Aris substituted or unsubstituted phenyl or monocyclic heteroaryl, the sumof m and o is from 1 to 3, and wherein one CH₂ may be substituted with Sor O.

In other words, while not intending to be limiting, A may be —(CH₂)₆—,cis —CH₂CH═CH—(CH₂)₃—, or —CH₂C≡C—(CH₂)₃—.

Alternatively, A may be a group which is related to one of these threemoieties in that any carbon is substituted with S or O. For example,while not intending to limit the scope of the invention in any way, Amay be an S substituted moiety such as one of the following or the like.

Alternatively, while not intending to limit the scope of the inventionin any way, A may be an O substituted moiety such as one of thefollowing or the like.

Alternatively, while not intending to limit the scope of the inventionin any way, A may have both an O and an S substituted in the chain, suchas one of the following or the like.

Alternatively, while not intending to limit the scope of the inventionin any way, in certain embodiments A is —(CH₂)_(m)—Ar—(CH₂)_(o)— whereinAr is substituted or unsubstituted phenyl or monocyclic heteroaryl, thesum of m and o is from 1 to 4, and wherein one CH₂ may be substitutedwith S or O. In other words, while not intending to limit the scope ofthe invention in any way, A comprises from 1 to 4 CH₂ moieties and Ar,e.g. —CH₂—Ar—, —(CH₂)₂—Ar—, —CH₂—ArCH₂—, —CH₂Ar(CH₂)₂—,—(CH₂)₂—Ar(CH₂)₂—, and the like; or

-   A comprises O, from 0 to 3 CH₂ moieties, and Ar, as in for example,    —O—Ar—, Ar—CH₂—O—, —O—Ar—(CH₂)₂—, —O—CH₂—Ar—, —O—CH₂—Ar—(CH₂)₂, and    the like; or-   A comprises S, from 0 to 3 CH₂ moieties, and Ar, as in for example,    —S—Ar—, Ar—CH₂—S—, —S—Ar—(CH₂)₂—, —S—CH₂—Ar—, —S—CH₂—Ar—(CH₂)₂, and    the like.

Ar is substituted or unsubstituted phenyl or substituted orunsubstituted monocyclic heteroaryl. In one embodiment, Ar issubstituted or unsubstituted phenyl, thienyl, furyl, or pyridinyl. Inanother embodiment Ar is phenyl (Ph). In another embodiment A is—(CH₂)₂—Ph. While not intending to limit scope of the invention in anyway, substituents may have 4 or less heavy atoms, or in other words, nonhydrogen atoms. Any number of hydrogen atoms required for a particularsubstituent will also be included. Thus, the substituent may be C4 orlower hydrocarbyl, including C4 or lower alkyl, alkenyl, alkynyl, andthe like; C3 or lower hydrocarbyloxy; CF₃; halo, such as F, Cl, or Br;hydroxyl; NH₂ and alkylamine functional groups up to C3; other N or Scontaining substituents; and the like.

In one embodiment A is —(CH₂)_(m)—Ar—(CH₂)_(o)— wherein Ar is phenyl,the sum of m and o is from 1 to 3, and wherein one CH₂ may besubstituted with S or O.

In another embodiment A is —CH₂—Ar—OCH₂—. In another embodiment A is—CH₂—Ar—OCH₂— and Ar is phenyl.

In another embodiment A is —(CH₂)₆—, cis —CH₂CH═CH—(CH₂)₃—, or—CH₂C≡C—(CH₂)₃—, wherein 1 or 2 carbon atoms may be substituted with Sor O; or A is —(CH₂)₂—Ph— wherein one CH₂ may be substituted with S orO.

In another embodiment A is —(CH₂)₆—, cis —CH₂CH═CH—(CH₂)₃—, or—CH₂C≡C—(CH₂)₃—, wherein 1 or 2 carbon atoms may be substituted with Sor O; or A is —(CH₂)₂—Ph—.

D is a moiety comprising from 1 to 6 non-hydrogen atoms, in other words,there are from 1 to 6 atoms which are not hydrogen, and any number ofhydrogen atoms required to form the complete substituent. For example, amethyl substituent has 1 carbon atom and 3 hydrogen atoms. Other examplesubstituents include other hydrocarbyl moieties comprising from 1 to 6carbon atoms including alkyl such as ethyl, propyl, isopropyl, butyl andisomers thereof, pentyl and isomers thereof, hexyl and isomers thereof,cyclic and unsaturated hydrocarbyls having 1 to 6 carbon atoms; CO₂H andsalts thereof, alkoxy up to C₅ such as methoxy, ethoxy, propoxy,isopropoxy, a butoxy isomer, or a pentoxy isomer; carboxylic acidesters; CN; NO₂; CF₃; F; Cl; Br; I; sulfonyl esters; SO₃H and saltsthereof, and the like. D may be in any reasonable position on the phenylring.

In certain compounds, n is 0. In other compounds n is 1, in othercompounds n is 2, and in other compounds n is 3.

A hydrocarbyl moiety refers to a moiety consisting of only carbon andhydrogen. While not intending to limit the scope of the invention in anyway, examples of different types of hydrocarbyl moiety are as follows.

On type of hydrocarbyl is alkyl including:

-   a) linear alkyl such as methyl, ethyl, propyl, n-butyl, n-pentyl,    n-hexyl, and the like;-   b) branched alkyl such as isopropyl, branched butyl isomers (i.e.    sec-butyl, tert-butyl, etc), branched pentyl isomers (i.e.    isopentyl, etc), branched hexyl isomers, and higher branched alkyl    fragments;-   c) cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl,    cyclohexyl, cycloheptyl, etc.; and-   d) alkyl fragments consisting of both cyclic and noncyclic    components, whether linear or branched, which may be attached to the    remainder of the molecule at any available position including    terminal, internal, or ring carbon atoms.

In analogy to alkyl, there is linear, branched, cycloalkyl, andcombination hydrocarbyl.

Another type of hydrocarbyl is alkenyl, which is similar to alkyl withthe exception that a double bond is present.

Another type of hydrocarbyl is alk(poly)enyl, which is similar toalkenyl, except that more than one double bond is present.

Another type of hydrocarbyl is alkynyl or an alk(poly)ynyl, which issimilar to alkenyl or alk(poly)ynyl except that one or more triple bondsare present.

Another type of hydrocarbyl is aryl, which includes phenyl, naphthyl andother aromatic hydrocarbyls.

Additionally, combinations of any of the above in any manner imaginableto those of ordinary skill in the art are also hydrocarbyl.

A hydrocarbyl moiety comprising a cyclic structure comprises acycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkyl(poly)enyl,cycloalkyl(poly)ynyl, aryl, and the like; and may consist of only thering or may be a combination of the ring and one or more of the linear,branched, or cyclic hydrocarbyl fragments; or may be a fused polycyclicstructure.

A hydroxyhydrocarbyl moiety consists of a combination of a hydrocarbylmoiety and a hydroxyl group. In other words, a hydrogen atom of thehydrocarbyl moiety is substituted with a hydroxyl group. Thehydroxyhydrocarbyl moiety attaches to the remainder of the molecule at acarbon atom.

Thus, while not intending to limit the scope of the invention in anyway, as R is a hydrocarbyl or a hydroxyhydrocarbyl moiety comprisingfrom 1 to 12 atoms, embodiments having R as any of the hydrocarbyl orhydroxycarbyl moieties listed above are specifically contemplatedherein. R may also be a different moiety which may be consideredhydrocarbyl or hydroxyhydrocarbyl according to the description givenherein.

In certain compounds, R is a hydroxyhydrocarbyl having the hydroxylgroup attached to the carbon atom which is also attached to theremainder of the molecule. In other words the hydroxyl group and theremainder of the molecule are on geminal positions on the hydrocarbylmoiety. This type of hydroxyhydrocarbyl moiety is referred to as a1-hydroxyhydrocarbyl moiety herein. Non-linear hydroxyhydrocarbyl ishydroxyhydrocarbyl wherein the hydrocarbyl portion is not linear, i.e.it has branching and/or a ring.

In other compounds R is hydroxyhydrocarbyl having the hydroxyl groupattached to a carbon atom which is directly attached to the remainingpart of the molecule. These particular hydroxyhydrocarbyl are called2-hydroxyhydrocarbyl herein. For example, —C(CH₃)₂CH₂OH is2-hydroxyhydrocarbyl. While not intending to limit the scope of theinvention in any way, a general structure where R is 2-hydrocarbyl isshown below.

As with all other structures shown herein, pharmaceutically acceptablesalts and prodrugs of compounds represent by these structures are alsocontemplated.

In one embodiment related to the above structure, R³, R⁴, and R⁵ areindependently H or C₁₋₆ alkyl. As the dashed line indicates the presenceor absence of a bond, R⁴ and R⁵ may be two separate moieties. Forexample, while not intending to be limiting, R⁴ and R⁵ may be methyl,and no bond would be present where indicated by the dashed line.Alternatively, while not intending to limit the scope of the inventionin any way, R⁴ and R⁵ may form a ring. In other words, a compound suchas the one shown below is possible, wherein x is from 1 to 6.

Pharmaceutically acceptable salts and prodrugs of compounds represent bythese structures are also contemplated.

In certain compounds, R comprises from 6 to 9 carbon atoms and a cyclicstructure. In other compounds, R comprises from 1 to 5 carbon atoms. Incertain compounds R is hydroxyalkyl having from 1 to 5 carbon atoms. Inother compounds R is a 1-hydroxyhydrocarbyl moiety comprising from 6 to9 carbon atoms and a cyclic structure. In other compounds R is a1-hydroxyhydrocarbyl moiety comprising from 6 to 9 carbon atoms and acyclic structure comprising from 4-7 carbon atoms. In other words, thecyclic structure part of R is a cyclobutyl, cyclopentyl, cyclohexyl, orcycloheptyl fragment. The cyclic structure part of R may also be acycloalkenyl or cycloalkynyl fragment such as cyclopentene orcyclohexene. In other compounds R is a hydrocarbyl moiety comprisingfrom 1 to 5 carbon atoms. In other words, R is methyl, ethyl, propyl,isopropyl, a butyl isomer such as t-butyl, or a pentyl isomer. Incertain compounds R is t-butyl.

Certain R groups are specifically contemplated herein. These are shownbelow, where PR represents the remaining part of the molecule.

As there is a dashed line between R and the phenyl ring, cyclicstructures having two carbon atoms of the phenyl ring are possible.While not intending to limit the scope of the invention in any way,compounds such as those represented by the structure below are thereforepossible.

Pharmaceutically acceptable salts and prodrugs thereof are alsocontemplated.

Other useful compounds comprise

or a pharmaceutically acceptable salt, or a prodrug thereof.

Other useful compounds comprise

or a pharmaceutically acceptable salt, or a prodrug thereof.

Those of ordinary skill in the art understand that any value whichrefers to the number of atoms, moieties, etc., on a small molecule willbe an integer, i.e. 0, 1, 2, 3, etc.

Certain useful compounds comprise

or a pharmaceutically acceptable salt, or a prodrug thereof.

Other useful compounds comprise

or a pharmaceutically acceptable salt, or a prodrug thereof.

Other useful examples of compounds comprise

or a pharmaceutically acceptable salt, or a prodrug thereof.

Other compounds comprise

or a pharmaceutically acceptable salt, or a prodrug thereof.

Other embodiments comprise

or a pharmaceutically acceptable salt, or a prodrug thereof,

-   -   wherein a dashed line indicates the presence or absence of a        bond.

Other compounds comprise

wherein X is C═O or CHCl; and

-   -   R is alkyl having from 3 to 6 carbon atoms.

Other compounds comprise

or a pharmaceutically acceptable salt, or a prodrug thereof,

-   -   wherein R⁶ is cycloalkyl comprising from 3 to 10 carbon atoms;        and    -   X is C═O or CHCl.

Other compounds comprise

or a pharmaceutically acceptable salt, or a prodrug thereof.

Other embodiments comprise

or a pharmaceutically acceptable salt, or a prodrug thereof

-   -   wherein q is an integer having a value of from 0 to 3.

Other compounds comprise

or a pharmaceutically acceptable salt, or a prodrug thereof.

Other compounds comprise

or a pharmaceutically acceptable salt, or a prodrug thereof.

Other useful compounds comprise

or a pharmaceutically acceptable salt, or a prodrug thereof.

Other useful embodiments comprise

or a pharmaceutically acceptable salt, or a prodrug thereof.

Another useful compound is

or a pharmaceutically acceptable salt, or a prodrug thereof.

Another useful compound is

or a pharmaceutically acceptable salt, or a prodrug thereof.

Another useful compound is

or a pharmaceutically acceptable salt, or a prodrug thereof.

Another useful compound is

or a pharmaceutically acceptable salt, or a prodrug thereof.

Another useful compound is

or a pharmaceutically acceptable salt, or a prodrug thereof.

Certain compounds comprise

or a pharmaceutically acceptable salt or a prodrug thereof.

Other compounds comprise

or a pharmaceutically acceptable salt or a prodrug thereof.

Another useful compound is

or a pharmaceutically acceptable salt, or a prodrug thereof.

Another useful compound is

or a pharmaceutically acceptable salt or a prodrug thereof.

Another useful compound is

or a pharmaceutically acceptable salt or a prodrug thereof.

Other compounds comprise

or a pharmaceutically acceptable salt or a prodrug thereof

-   -   wherein X is C═O or CHCl.        Another useful compound is

or a pharmaceutically acceptable salt or a prodrug thereof.

A prostaglandin EP₂ selective agonist is a compound which is more activeat a prostaglandin EP₂ receptor than at any other prostaglandinreceptor.

In one embodiment, the compound has an IC₅₀ value less than 1 μM. Inanother embodiment, the compound is more than 100 times more active atthe EP₂ receptor than at any other receptor. In another embodiment, thecompound is more than 1000 times more active at the EP₂ receptor than atany other receptor.

The ω-chain has the meaning normally understood in the art. Inprostaglandin E₂, the ω-chain is in the third position of thecyclopentanone ring, where the position 1 is the carbonyl and theα-chain is at position 2. However, the meaning of the term α-chainshould be adapted according to synthetic variations that are made toprostaglandin E₂. A person of ordinary skill in the art can readilydiscern the ω-chain in synthetic analogs and derivatives ofprostaglandin E₂. For example, while not intending to limit the scope ofthe invention in any way, the ω-chain could be at the third position ina 1-chlorocyclopentane having the α-chain in the 2 position.

A substituted phenyl, wherein at least one substituent consists ofhydrocarbyl or non-linear hydroxyhydrocarbyl may have additionalsubstituents which are not hydrocarbyl or non-linear hydroxyhydrocarbyl,i.e. at least one substituent is hydrocarbyl or non-linearhydroxyhydrocarbyl and at least one substituent is not.

The compounds of disclosed herein are useful for the prevention ortreatment of glaucoma or ocular hypertension in mammals, or for themanufacture of a medicament for the treatment of glaucoma or ocularhypertension. They are also useful for the treatment of those diseasesdisclosed in the art as being amenable to treatment by prostaglandin EP₂agonist, such as the ones listed previously.

A number of exemplary compounds, and exemplary methods of making thesecompounds, are identified in the following patent applications, all ofwhich are expressly incorporated by reference herein:

U.S. Provisional Patent Application No. 60/678,403, filed on May 6,2005;

U.S. Provisional Patent Application No. 60/742,779, filed on Dec. 6,2005;

U.S. Provisional Patent Application No. 60/777,506, filed Feb. 29, 2006;

U.S. Provisional Patent Application No. 60/660,748, filed on Mar. 10,2005;

U.S. Provisional Patent Application No. 60/783,979; filed on Mar. 20,2005;

U.S. Provisional Patent Application No. 60/744,236; filed on Apr. 4,2006;

U.S. Provisional Patent Application No. 60/746,393; filed on May 4,2006;

U.S. Provisional Patent Application No. 60/746,391; filed on May 4,2006;

U.S. Provisional Patent Application No. 60/747,835; filed on May 22,2006;

U.S. Provisional Patent Application No. 60/747,115; filed on May 2,2006; and

U.S. Provisional Patent Application No. 60/803,040, filed on May 24,2006.

Those skilled in the art will readily understand that for administrationor the manufacture of medicaments the compounds disclosed herein can beadmixed with pharmaceutically acceptable excipients which per se arewell known in the art. Specifically, a drug to be administeredsystemically, it may be confected as a powder, pill, tablet or the like,or as a solution, emulsion, suspension, aerosol, syrup or elixirsuitable for oral or parenteral administration or inhalation.

For solid dosage forms or medicaments, non-toxic solid carriers include,but are not limited to, pharmaceutical grades of mannitol, lactose,starch, magnesium stearate, sodium saccharin, the polyalkylene glycols,talcum, cellulose, glucose, sucrose and magnesium carbonate. The soliddosage forms may be uncoated or they may be coated by known techniquesto delay disintegration and absorption in the gastrointestinal tract andthereby provide a sustained action over a longer period. For example, atime delay material such as glyceryl monostearate or glyceryl distcaratemay be employed. They may also be coated by the technique described inthe U.S. Pat. Nos. 4,256,108; 4,166,452; and 4,265,874 to form osmotictherapeutic tablets for control release. Liquid pharmaceuticallyadministrable dosage forms can, for example, comprise a solution orsuspension of one or more of the presently useful compounds and optionalpharmaceutical adjutants in a carrier, such as for example, water,saline, aqueous dextrose, glycerol, ethanol and the like, to therebyform a solution or suspension. If desired, the pharmaceuticalcomposition to be administered may also contain minor amounts ofnontoxic auxiliary substances such as wetting or emulsifying agents, pHbuffering agents and the like. Typical examples of such auxiliary agentsare sodium acetate, sorbitan monolaurate, triethanolamine, sodiumacetate, triethanolamine oleate, etc. Actual methods of preparing suchdosage forms are known, or will be apparent, to those skilled in thisart; for example, see Remington's Pharmaceutical Sciences, MackPublishing Company, Easton, Pa., 16th Edition, 1980. The composition ofthe formulation to be administered, in any event, contains a quantity ofone or more of the presently useful compounds in an amount effective toprovide the desired therapeutic effect.

Parenteral administration is generally characterized by injection,either subcutaneously, intramuscularly or intravenously. Injectables canbe prepared in conventional forms, either as liquid solutions orsuspensions, solid forms suitable for solution or suspension in liquidprior to injection, or as emulsions. Suitable excipients are, forexample, water, saline, dextrose, glycerol, ethanol and the like. Inaddition, if desired, the injectable pharmaceutical compositions to beadministered may also contain minor amounts of non-toxic auxiliarysubstances such as wetting or emulsifying agents, pH buffering agentsand the like.

The amount of the presently useful compound or compounds administeredis, of course, dependent on the therapeutic effect or effects desired,on the specific mammal being treated, on the severity and nature of themammal's condition, on the manner of administration, on the potency andpharmacodynamics of the particular compound or compounds employed, andon the judgment of the prescribing physician. The therapeuticallyeffective dosage of the presently useful compound or compounds ispreferably in the range of about 0.5 or about 1 to about 100 mg/kg/day.

A liquid which is ophthalmically acceptable is formulated such that itcan be administered topically to the eye. The comfort should bemaximized as much as possible, although sometimes formulationconsiderations (e.g. drug stability) may necessitate less than optimalcomfort. In the case that comfort cannot be maximized, the liquid shouldbe formulated such that the liquid is tolerable to the patient fortopical ophthalmic use. Additionally, an ophthalmically acceptableliquid should either be packaged for single use, or contain apreservative to prevent contamination over multiple uses.

For ophthalmic application, solutions or medicaments are often preparedusing a physiological saline solution as a major vehicle. Ophthalmicsolutions should preferably be maintained at a comfortable pH with anappropriate buffer system. The formulations may also containconventional, pharmaceutically acceptable preservatives, stabilizers andsurfactants.

Preservatives that may be used in the pharmaceutical compositions of thepresent invention include, but are not limited to, benzalkoniumchloride, chlorobutanol, thimerosal, phenylmercuric acetate andphenylmercuric nitrate. A useful surfactant is, for example, Tween 80.Likewise, various useful vehicles may be used in the ophthalmicpreparations of the present invention. These vehicles include, but arenot limited to, polyvinyl alcohol, povidone, hydroxypropyl methylcellulose, poloxamers, carboxymethyl cellulose, hydroxyethyl celluloseand purified water.

Tonicity adjustors may be added as needed or convenient. They include,but are not limited to, salts, particularly sodium chloride, potassiumchloride, mannitol and glycerin, or any other suitable ophthalmicallyacceptable tonicity adjustor.

Various buffers and means for adjusting pH may be used so long as theresulting preparation is ophthalmically acceptable. Accordingly, buffersinclude acetate buffers, citrate buffers, phosphate buffers and boratebuffers. Acids or bases may be used to adjust the pH of theseformulations as needed.

In a similar vein, an ophthalmically acceptable antioxidant for use inthe present invention includes, but is not limited to, sodiummetabisulfite, sodium thiosulfate, acetylcysteine, butylatedhydroxyanisole and butylated hydroxytoluene.

Other excipient components which may be included in the ophthalmicpreparations are chelating agents. A useful chelating agent is edetatedisodium, although other chelating agents may also be used in place orin conjunction with it.

The ingredients are usually used in the following amounts:

Ingredient Amount (% w/v) active ingredient about 0.001-5 preservative  0-0.10 vehicle 0-40 tonicity adjustor 1-10 buffer 0.01-10   pHadjustor q.s. pH 4.5-7.5 antioxidant as needed surfactant as neededpurified water as needed to make 100%

For topical use, creams, ointments, gels, solutions or suspensions,etc., containing the compound disclosed herein are employed. Topicalformulations may generally be comprised of a pharmaceutical carrier,cosolvent, emulsifier, penetration enhancer, preservative system, andemollient.

The actual dose of the active compounds of the present invention dependson the specific compound, and on the condition to be treated; theselection of the appropriate dose is well within the knowledge of theskilled artisan.

EXAMPLE 2

Binding Data

Ki

Competition binding experiments were performed in a medium containingHank's balanced salt solution, Hepes 20 mM, pH 7.3, membranes (˜60 μgprotein) or 2×10⁵ cells from HEK 293 cells stably expressing human EP2receptors, [³H]PGE2 (10 nM) and various concentrations of test compoundsin a total volume of 300 μl. Reaction mixtures were incubated at 23° C.for 60 min, and were filtered over Whatman GF/B filters under vacuum.Filters were washed three times with 5 ml ice-cold buffer containing 50mM Tris/HCl (pH 7.3). Non-specific binding was estimated in the presenceof excess unlabeled PGE2 (10 μM). Binding data fitted to the bindingmodel for a single class of binding sites, using nonlinear regressionanalysis. IC₅₀ values thus obtained were converted to Ki using theequation of Ki=(IC₅₀/(1+[L]/K_(D)) where [L] represents PGE2concentration (10 nM) and K_(D) the dissociation constant for [³H]PGE2at human EP2 receptors (40 nM).

Radioligand Binding

Cells Stably Expressing EP₁, EP₂, EP₄ and FP Recptors

HEK-293 cells stably expressing the human or feline FP receptor, or EP₁,EP₂, or EP₄ receptors were washed with TME buffer, scraped from thebottom of the flasks, and homogenized for 30 sec using a Brinkman PT10/35 polytron. TME buffer was added to achieve a final 40 ml volume inthe centrifuge tubes (the composition of TME is 100 mM TRIS base, 20 mMMgCl₂, 2M EDTA; 10N HCl is added to achieve a pH of 7.4).

The cell homogenate was centrifuged at 19000 r.p.m. for 20 min at 4° C.using a Beckman Ti-60 rotor. The resultant pellet was resuspended in TMEbuffer to give a final 1 mg/ml protein concentration, as determined byBiorad assay. Radioligand binding competition assays vs. [³H-]17 —phenylPGF_(2α) (5 nM) were performed in a 100 μl volume for 60 min. Bindingreactions were started by adding plasma membrane fraction. The reactionwas terminated by the addition of 4 ml ice-cold TRIS-HCl buffer andrapid filtration through glass fiber GF/B filters using a Brandel cellharvester. The filters were washed 3 times with ice-cold buffer and ovendried for one hour.

[³H-] PGE₂ (specific activity 180 Ci mmol) was used as the radioligandfor EP receptors. [³H] 17-phenyl PGF_(2α) was employed for FP receptorbinding studies. Binding studies employing EP₁, EP₂, EP₄ and FPreceptors were performed in duplicate in at least three separateexperiments. A 200 μl assay volume was used. Incubations were for 60 minat 25° C. and were terminated by the addition of 4 ml of ice-cold 50 mMTRIS-HCl, followed by rapid filtration through Whatman GF/B filters andthree additional 4 ml washes in a cell harvester (Brandel). Competitionstudies were performed using a final concentration of 5 nM [³H]—PGE₂, or5 nM [³H] 17-phenyl PGF_(2α) and non-specific binding determined with10⁻⁵M of unlabeled PGE₂, or 17-phenyl PGF_(2α), according to receptorsubtype studied.

Methods for FLIPR™ Studies

-   (a) Cell Culture

HEK-293(EBNA) cells, stably expressing one type or subtype ofrecombinant human prostaglandin receptors (prostaglandin receptorsexpressed: hDP/Gqs5; hEP₁; hEP₂/Gqs5; hEP_(3A)/Gqi5; hEP₄/Gqs5; hFP;hIP; hTP), were cultured in 100 mm culture dishes in high-glucose DMEMmedium containing 10% fetal bovine serum, 2 mM 1-glutamine, 250 μg/mlgeneticin (G418) and 200 μg/ml hygromycin B as selection markers, and100 units/ml penicillin G, 100 μg/ml streptomycin and 0.25 μg/mlamphotericin B.

-   (b) Calcium Signal Studies on the FLIPR™

Cells were seeded at a density of 5×10⁴ cells per well in Biocoat®Poly-D-lysine-coated black-wall, clear-bottom 96-well plates(Becton-Dickinson) and allowed to attach overnight in an incubator at37° C. Cells were then washed two times with HBSS-HEPES buffer (HanksBalanced Salt Solution without bicarbonate and phenol red, 20 mM HEPES,pH 7.4) using a Denley Cellwash plate washer (Labsystems). After 45minutes of dye-loading in the dark, using the calcium-sensitive dyeFluo-4 AM at a final concentration of 2 μM, plates were washed fourtimes with HBSS-HEPES buffer to remove excess dye leaving 100 μl in eachwell. Plates were re-equilibrated to 37° C. for a few minutes.

Cells were excited with an Argon laser at 488 nm, and emission wasmeasured through a 510-570 nm bandwidth emission filter (FLIPR™,Molecular Devices, Sunnyvale, Calif.). Drug solution was added in a 50μl volume to each well to give the desired final concentration. The peakincrease in fluorescence intensity was recorded for each well. On eachplate, four wells each served as negative (HBSS-HEPES buffer) andpositive controls (standard agonists: BW245C (hDP); PGE₂ (hEP₁;hEP₂/Gqs5; hEP_(3A)/Gqi5; hEP₄/Gqs5); PGF_(2α) (hFP); carbacyclin (hIP);U-46619 (hTP), depending on receptor). The peak fluorescence change ineach drug-containing well was then expressed relative to the controls.

Compounds were tested in a high-throughput (HTS) orconcentration-response (CoRe) format. In the HTS format, forty-fourcompounds per plate were examined in duplicates at a concentration of10⁻⁵ M. To generate concentration-response curves, four compounds perplate were tested in duplicates in a concentration range between 10⁻⁵and 10⁻¹¹ M. The duplicate values were averaged. In either, HTS or CoReformat each compound was tested on at least 3 separate plates usingcells from different passages to give an n≧3.

The results of the binding and activity studies, presented in the Tablesbelow, demonstrate that the compounds disclosed herein are selectiveprostaglandin EP₂ agonists, and are thus useful for the treatment ofglaucoma, ocular hypertension, the other diseases or conditionsdisclosed herein.

BINDING IC50 (nm) FUNCTIONAL EC50 (nm) STRUCTURE HEP2 HEP3 HEP4 HFP HEP1HEP2 HEP3A HEP4 HTP HIP HDP

NA NA >10 K >10 K >10 K NA NA NA

NA NA >10 K >10 K >10 K NA >10 K >10 K

NA NA 5294 1698 NA NA NA NA

NA NA 5259 NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA >10 K NA >10 K NA NA NA

NA NA NA NA NA NA NA NA

NA NA >10 K >10 K NA >10 K NA NA

NA NA 322 455 NA >10 K NA >10 K

NA NA NA NA NA NA NA NA

NA NA 1479 3118 NA NA NA NA

NA NA NA NA NA >10 K NA NA

NA NA >10 K NA NA NA NA NA

NA NA >10 K NA NA NA NA NA

NA NA 3723 NA NA NA NA NA

NA NA 635 NA NA NA NA NA

NA NA 2270 NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA 546 NA NA NA NA NA

NA NA >10 K NA NA >10 K NA NA

NA >10 K 1709 NA NA NA NA NA

NA NA 936 >10 K >10 K >10 K NA NA

NA NA >10 K >10 K NA NA NA NA

NA >10 K 102 3390 NA 4273 >10 K NA

NA >10 K 118 2053 >10 K 1269 NA >10 K

NA NA >10 K NA NA NA NA NA

NA >10 K 264 >10 K NA >10 K NA >10 K

NA NA NA NA NA NA NA

NA NA >10 K NA >10 K NA NA NA

NA NA >10 K NA NA NA NA NA

NA NA >10 K NA NA NA NA NA

NA NA 450 NA NA NA NA NA

NA NA >10 K NA NA NA NA NA

NA NA 392 NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA 3445 NA NA NA NA >10 K

NA NA 2813 NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA >10 K

NA NA NA NA NA NA NA NA

5200 NA NA NA NA 266 NA NA NA NA

NA NA 3844 NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA 20773 NA NA NA NA NA

NA NA 1550 NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA

NA NA >10 K >10 K >10 K >10 K NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA >10 K NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA

2281 NA NA 405 NA NA NA NA NA

13139 NA NA 529 NA NA 2993 NA NA

NA NA NA NA 22457 17525 NA NA

13100 NA NA NA NA NA 506 NA NA

6251 NA 221 818 NA NA 200 NA

NA NA NA NA NA NA NA NA

NA NA >10 K NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA >10 K NA NA NA >10 K NA

NA NA >10 K NA NA NA NA NA

NA NA 513 NA NA >10 K NA NA

NA NA >10 K NA NA >10 K NA NA

NA NA 743 NA NA >10 K NA NA

NA >10 K

NA NA >10 K NA NA NA NA

NA NA >10 K NA 26289 NA NA NA

18735 NA NA 526 NA NA NA NA

48765 >10 K >10 K

4185 NA NA 173 NA NA NA NA

>10 K >10 K

8950 NA NA 708 NA >10 K NA NA NA

NA NA

NA NA

NA NA >10 K NA NA NA NA NA

NA 1873 3128 NA NA 94 NA NA

NA NA

NA NA

>10 K NA

13150 NA NA NA NA NA NA NA

NA >10 K

>10 K NA

NA NA 4995 NA >10 K NA NA NA

1537 NA NA 779 NA >10 K NA NA NA

NA NA 6575 NA >10 K NA NA NA

3630 NA NA 1265 NA >10 K NA NA NA

>10 K >10 K

Ki 1340 NA NA 497 NA NA NA NA NA

>10 K >10 K

Ki 3022 >10 K NA

EP2 data Ki EP4 data flipr cAMP pH flipr Other Receptors (EC50 in nM)Structure EC50 EC50 Ki 6.0 EC50 KI hFP hEP1 hEP3A hTP hIP hDP

>10000 517 NA NA >10000 NA NA NA NA NA NA

212 8 387 NA >10000 NA NA NA NA NA NA

20 1.5 190 NA >10000 NA NA NA NA NA 5763

426 27 1639 NA >10000 NA NA NA NA NA NA

1812 312 5731 >10000 >10000 NA NA NA NA NA 7560

226 15 1382 NA >10000 NA NA NA NA NA NA

5 0.55 23 NA >10000 NA NA 155 NA NA 1234

16 1.6 31 >10000 >10000 NA NA 2345 NA NA 7695

215 8 163 NA >10000 NA NA NA NA NA NA

62 5 345 >10000 >10000 NA NA 153 NA NA 7749

15 1.5 116 >10000 6032 NA NA 1205 NA NA 6800

6 0.19 21 >10000 >10000 NA NA 12 NA NA 812

1.6 0.15 15 >10000 4849 NA NA 156 NA NA 296

134 7 229 NA 3842 NA NA 71 NA NA 6829

49 4 201 NA 3288 NA NA 621 NA NA NA

30 0.9 10 >10000 NA NA 47 NA NA 105

16 1.4 12 NA NA 6952 7 NA NA 37

5 0.6 8 NA NA NA 33 NA >10000 106

728 NA 22726 NA 8752 NA NA NA NA NA NA

>10000 NA >10000 >10000

>10000 NA >10000 >10000

442 28 4000 189 >10000 >10000 NA NA >10000 NA NA 1921

1343 51 501 27 >10000 >10000 NA >10000 19234 >10000 NA 2323

4121 548 >10000 >10000 >10000 NA >10000 19544 NA NA >10000

388 26 2028 NA >10000 NA NA 1927 NA NA NA

7669 1218 >10000 NA >10000 NA NA NA NA NA NA

1228 148 2293 NA >10000 NA NA NA NA NA NA

8 3 115 >10000 >10000 NA NA 403 NA NA 3233

>10000 517 NA NA >10000 NA NA NA NA NA NA

212 8 387 NA >10000 NA NA NA NA NA NA

20 1.5 190 NA >10000 NA NA 519 NA NA 5763

426 27 1639 NA >10000 NA NA NA NA NA NA

1812 312 5731 >10000 >10000 NA NA NA NA NA 7560

226 15 1382 NA >10000 NA NA 1411 NA NA NA

5 0.55 23 NA >10000 NA NA 155 NA NA 1234

16 1.6 31 >10000 >10000 NA NA 2345 NA NA 7695

215 8 163 NA >10000 NA NA NA NA NA NA

62 5 345 >10000 >10000 NA NA 153 NA NA 7749

15 1.5 116 >10000 6032 NA NA 1205 NA NA 6800

6 0.19 21 >10000 >10000 NA NA 12 NA NA 812

1.6 0.15 15 >10000 4849 NA NA 156 NA NA 296

134 7 229 NA 3842 NA NA 71 NA NA 6829

49 4 201 NA 3288 NA NA 621 NA NA NA

30 0.9 10 >10000 NA NA 47 NA NA 105

16 1.4 12 NA NA 6952 7 NA NA 37

5 0.6 8 NA NA NA 33 NA >10000 106

7284 NA 22726 NA 8752 NA NA NA NA NA NA

FUNCTIONAL (Ca2+, EC50, nM) BINDING (K_(i), nM) HE HE HE HE HF HE HE P3HE HT HD STRUCTURE P2 P3 P4 P P1 P2 A P4 P HIP P

NA NA NA NA NA >10 K NA NA

NA NA 265 7 278 9 NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA 340 7 382 8 NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA 287 2 NA NA NA NA >10 K

NA NA NA NA NA NA NA NA

NA NA 105 0 NA NA NA NA NA

NA NA >10 K NA >10 K NA NA NA

NA NA >10 K NA NA NA NA NA

NA NA 164 0 NA NA NA NA NA

NA NA >10 K NA NA >10 K NA NA

NA NA 204 7 NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA >10 K NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA 210 1 NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA >10 K NA NA >10 K NA NA

54 NA NA 2 (3)^(a) 55 >10 K NA NA NA

TABLE 2 BINDING FUNCTIONAL STRUCTURE HEP2 HEP2

15000 15000

700 1011

4600 15000

700 343

50000 50000

15000 50000

50000 50000

600 15000

50000 50000

1900 15000

1400 1409

50000 50000

15000 15000

7100 7100

600 769

50000 50000

2811 4877

TABLE 3 BINDING (K₁, nM) FUNCTIONAL (Ca2+, EC50, nM) STRUCTURE HEP2 HEP3HEP4 HFP HEP1 HEP2 HEP3A HEP4 HTP HIP HDP

4677 NA 15000 NA NA 2162 3090 NA NA NA NA

871 5200 15000 NA NA 284 90 NA NA NA NA

15000 15000 NA NA 15000 15000 NA NA NA NA

700 15000 15000 NA NA 1011 552 NA NA NA NA EP2 EP4 OTHER cAMP Ca2+ cAMPKi Ca2+ RECEPTORS EC50 Ki EC50 EC50 EC50 EC50 Ca2+ STRUCTURE (nM) (nM)(nM) (nM) (nM) (nM) EC50 (nM)

9 111 31 2809 NA NA: EP1, EP3, DP, FP, IP, TP BINDING -Ki (nM)Ca²⁺Signal-EC50 (nM)^(b) STRUCTURE^(a) EP2 EP4 FP EP1 EP2 EP3 EP4 TP IPDP

504 2364 not active not active 427 (58) 449 10,000 not active not activenot active

25 1400 not active not active 15 (4) 25 not active not active not activenot active

1252 >10K not active not active 34 (4) 10000 not active not active notactive not active (a) Refers to EC₅₀ in a cAMP mediated assay NA means“not active.”

In Vivo Testing

Intraocular Pressure (IOP)

Intraocular pressure studies in dogs involved pneumatonometry performedin conscious, Beagle dogs of both sexes (10-15 kg). The animals remainedconscious throughout the study and were gently restrained by hand. Drugswere administered topically to one eye as a 25 μL volume drop, the othereye received 25 μL vehicle (0.1% polysorbate 80:10 mM TRIS) as acontrol. Proparacaine (0.1%) was used for corneal anesthesia duringtonometry. Intraocular pressure was determined just before drugadministration and at 2, 4 and 6 hr thereafter on each day of the 5 daystudy. Drug was administered immediately after the first IOP reading.

Ocular Surface Hyperemia

Ocular surface hyperemia was visually assessed and scored according to asystem typically used clinically.

Hyperemia Score Assigned Value <1 trace 0.5 1 mild 1 moderate 2 severe 3

Ocular surface hyperemia was evaluated at the same time points asintraocular pressure measurement. It should be noted that untreated dogeyes frequently have a pink/red tone. Thus, values of trace or even mildare not necessarily out of the normal range. Similar tests were used todetermine ocular surface hyperemia on monkeys and rabbits. The Tablebelow shows results of testing for certain compounds.

DOG MONKEY Max. Max. RABBIT Conc. ΔIOP Max. ΔIOP Max. STRUCTURE (g/100mL) (mm Hg) hyperemia (mm Hg) hyperemia

0.10% −4 0.9 0

0.01% 8 0.1

0.10% −4 1 0

0.01% −4 0.6 5

0.30% −7 1 7

0.10% −12 0.25

0.30% −8 0.9

0.10% −5 0.7 6 0.1

0.1% −50 2.0 31

0.1% −37 0.8 20 0

0.1% −12 0.6 19 0.0

The foregoing description details specific methods and compositions thatcan be employed to practice the present invention, and represents thebest mode contemplated. However, it is apparent for one of ordinaryskill in the art that further compounds with the desired pharmacologicalproperties can be prepared in an analogous manner, and that thedisclosed compounds can also be obtained from different startingcompounds via different chemical reactions. Similarly, differentpharmaceutical compositions may be prepared and used with substantiallythe same result. Thus, however detailed the foregoing may appear intext, it should not be construed as limiting the overall scope hereof,rather, the ambit of the present invention is to be governed only by thelawful construction of the appended claims.

1. A compound having a formula

or a pharmaceutically acceptable salt or tautomer thereof; wherein thecompound has an EC₅₀ value of 20 nM or less in HEK-EBNA cells expressinga PG EP2 receptor subtype according to a cAMP assay.
 2. The compound ofclaim 1, wherein the compound reduces intraocular pressure of a beagledog having a mass of from about 10 to about 15 kg by at least 4 mmHgwhen administered topically in a liquid composition to an eye of saiddog, wherein said composition has concentration of about 0.3% (w/v). 3.The compound of claim 2 wherein the mass of the beagle dog is about 10kg.
 4. The compound of claim 2 wherein the mass of the beagle dog isabout 15 kg.
 5. The compound of claim 2 wherein intraocular pressure isobtained by pneumatonometry.
 6. The compound of claim 3 whereinintraocular pressure is obtained by pneumatonometry.
 7. The compound ofclaim 5 wherein the compound reduces the intraocular pressure of the dogby at least 4 mmHg at one or more of about 2, about 4, or about 6 hoursafter a single administration.
 8. The compound of claim 6 wherein thecompound reduces the intraocular pressure of the dog by at least 4 mmHgat one or more of about 2, about 4, or about 6 hours after a singleadministration.
 9. The compound of claim 8 wherein the compound reducesthe intraocular pressure of the dog by at least 4 mmHg at about 2 hoursafter a single administration.
 10. The compound of claim 8 wherein thecompound reduces the intraocular pressure of the dog by at least 4 mmHgat about 4 hours after a single administration.
 11. The compound ofclaim 8 wherein the compound reduces the intraocular pressure of the dogby at least 4 mmHg at about 6 hours after a single administration. 12.The compound of claim 8 wherein said liquid composition consists of thecompound having a concentration of about 0.3% (w/v) or less, 0.1% (w/v)polysorbate 80, 10 mM TRIS, and water.
 13. The compound of claim 9wherein said liquid composition consists of the compound having aconcentration of about 0.3% (w/v) or less, 0.1% (w/v) polysorbate 80, 10mM TRIS, and water.
 14. The compound of claim 2 wherein 0.1% (w/v)proparcaine is used for corneal anesthesia during pneumnatonometry. 15.The compound of claim 12 wherein 0.1% (w/v) proparcaine is used forcorneal anesthesia during pneumatonometry.
 16. The compound of claim 13wherein 0.1% (w/v) proparcaine is used for corneal anesthesia duringpneumatonometry.
 17. The compound of claim 2 wherein the compoundreduces the intraocular pressure of the dog by at least 4 mmHg at one ormore of about 2, about 4, or about 6 hours after a singleadministration.
 18. The compound of claim 2 wherein said liquidcomposition consists of the compound having a concentration of about0.3% (w/v) or less, 0.1% (w/v) polysorbate 80, 10 mM TRIS, and water.19. The compound of claim 1, wherein the compound reduces intraocularpressure of a human with glaucoma or elevated intraocular pressure by atleast 5 mmHg when administered topically in a liquid composition to aneye of said human, wherein compound has a concentration of about 0.3%(w/v) or less in said composition.